Test pLsrA/cI
Test pLsrA/cI promoter
The pLsrA/cI promoter is constitutively off and is repressed by the lsrR protein. The addition of AI-2 should de-repress the lsrR operon causing it to glow. The addition of the cI protein should repress the promoter, causing it to not glow.
Test pLsrA/cI repression:
pBAD→RBS+LsrR+TT
plsrA/cI→GFP does not glow
I13453+B0034+K091001+B0015+K091106+E0240
add AI-2
pBAD→RBS+LsrR+TT
plsrA/cI→GFP glows
I13453+B0034+K091001+B0015+K091106+E0240
LsrR | cI | AI-2 + GFP | AI-2 - GFP |
---|---|---|---|
0 | 0 | 1 | 1 |
1 | 0 | 1 | 0 |
0 | 1 | 0 | 0 |
1 | 1 | 0 | 0 |
Ligation plan for test sequences
K091106 ligated with E0240
need to ligate some promoter (digest with E/S) with an RBS (digest E/X) and ligate lsrR(digest with E/S) with a double terminator (digest with E/X)
then ligate the promoter+RBS to the 5' end of the lsrR+TT parts together
then ligate that part 5' to the plsrA/cI+GFP